The hypothesis of this proposal is that physiologic changes associated with stress, down-regulate pro-inflammatory cytokine, chemokine and growth factor gene expression resulting in alterations in cellular trafficking and activation, thus impairing wound healing. The long term goal of these studies is to understand the mechanisms underlying stress-related changes between the neuroendocrine and inflammatory responses that are responsible for altered wound healing. The aims of the proposal are 1) to determine the influence of stress on the pattern and kinetics of chemokine, pro-inflammatory cytokine and growth factor gene expression during the early stages of wound healing, 2) to determine the stress-induced, neuro-endocrine mechanisms that regulate pro-inflammatory cytokine, chemokine and growth factor gene expression during wound healing and 3) to determine the mechanism of the anti-glucocorticoid actions of androstenediol (a metabolite of DHEA) and delineate its ability to regulate pro-inflammatory cytokines (IL-1 alpha and beta and TNF alpha), chemokines (KC, IP10, MCP-1 and MIP 1 alpha) and growth factor (KGF, VEGF and TGF beta )gene expression as a therapeutic strategy to improve wound healing in stressed individuals. The proposal makes use of histological, molecular biology and pharmacological approaches to address the specific aims. The in vivo study of wound healing is accompanied by in vitro studies of the direct effect of glucocorticoids and AED (a metabolite of dihydroxyepiandrosterone) on cellular expression of transcription factors and cytokines. Restraint stress repeated over several days is to be used as a stress paradigm, skin wounding will be used to determine the rate of healing and the profile of cytokines and chemokines during the healing period. Subsequent studies will use an in vitro approach in which neutrophils and macrophages isolated from female mice will be incubated with corticosterone to determine the effect of LPS induced stimulation of cytokine and chemokine release and on changes in NF-kappa beta. In addition, the influence of AED on wound healing, cytokine and chemokine production and transcription factor NF-kappa beta will also be addressed in vivo.